A kind of Bacillus subtilis for preventing and controlling poultry enteritis and its application

A technology of Bacillus subtilis and Bacillus licheniformis, applied in the application, Lactobacillus, bacteria used in food preparation, etc., can solve the problems of gap in anti-infection effect, slow onset of microecological preparations and continuous use for a long time, and achieve improvement. Fecal status, reduced intestinal damage, and improved productivity

Active Publication Date: 2022-08-05
山东康地恩生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because probiotics can only partially replace antibiotics, compared with sensitive antibacterials, their anti-infection effects still have a certain gap
In addition, the probiotics have the disadvantages of slow onset and long-term continuous use during use.

Method used

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  • A kind of Bacillus subtilis for preventing and controlling poultry enteritis and its application
  • A kind of Bacillus subtilis for preventing and controlling poultry enteritis and its application
  • A kind of Bacillus subtilis for preventing and controlling poultry enteritis and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Strain isolation and screening

[0023] 1. Sample source: Intestinal feces of broilers from Gaomi farms in Weifang City, Shandong Province.

[0024] 2. Bacterial isolation and screening:

[0025] Weigh 1 g of fecal sample, put it in 9 mL of sterile physiological saline, and process it at 30°C, 200 rpm for 10 minutes to obtain a bacterial suspension; place the bacterial suspension in a water bath at 80°C for 10 minutes, take it out, and cool it quickly; The treated bacterial suspension was diluted to an appropriate concentration by a 10-fold gradient, 100uL was taken on the nutrient agar plate, smeared evenly, and cultivated at 37°C for 16-24 hours; pick out different single colonies that had been cultivated on the nutrient agar plate, and further streak 2 times, and purified to obtain a single strain. The applicant selected 5 strains of single bacteria and named them A, B, C, D, and E respectively.

[0026] The sporozoite liquid of Eimeria tenella was mixed...

Embodiment 2

[0031] Example 2 Identification of VB236 strain

[0032] 2.1 Identification of colony morphology

[0033] like figure 1 As shown, the colonies of the VB236 strain on the nutrient agar medium were yellowish, the surface was rough and opaque, and there was no mucus.

[0034] 2.2 16S rRNA molecular identification

[0035] The genome of the VB236 strain was extracted using a kit. The 16S rRNA was then amplified using specific primers using the genome as a template. The amplified PCR products were detected by 1% agarose gel electrophoresis and sent to a sequencing company for sequencing.

[0036] The sequencing results showed that the sequence of the PCR amplification product was SEQ ID NO: 1. By BLAST alignment of this sequence in the NCBI database, it was found that it is compatible with Bacillus subtilis ( Bacillus subtilis ) have the highest similarity. Therefore, the VB236 strain was initially identified as Bacillus subtilis ( Bacillus subtilis ).

[0037] 2.3 MALDI-...

Embodiment 3

[0052] Example 3 Evaluation of the anti-coccidial ability of Bacillus subtilis VB236

[0053] 1. Preparation of Eimeria tenella sporozoites

[0054] Transfer the purified Eimeria tenella sporulated oocyst suspension into a sterilized homogenizer for grinding to release the sporocysts, transfer the ground suspension into a conical flask, add 0.25% trypsin, 5% fresh chicken bile, add sterile PBS to 20 mL, and incubate in a 41°C water bath for 30 min with shaking. When 95% of the sporozoites overflowed by microscopy, the supernatant was removed by centrifugation, and washed repeatedly with PBS to remove bile and trypsin. Dilute to 0.5×10 with serum-containing, double-antibody DMEM medium 4 spare.

[0055] 2. Preparation of metabolites of test strain Bacillus subtilis VB 236

[0056] Bacillus subtilis VB 236 was inoculated into nutrient agar liquid medium, cultured with shaking at 37 °C for 56 h, the number of spores was observed by microscopy, centrifuged at 1000 r / min for 5 ...

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Abstract

The invention relates to the technical field of functional microorganism screening and application, and provides a Bacillus subtilis for preventing and controlling poultry enteritis and its application. The Bacillus subtilis has been deposited in the China Center for Type Culture Collection of Wuhan University, Wuhan, China on June 6, 2019, and the deposit number is CCTCC NO: M2019437. The Bacillus subtilis has a good preventive effect on intestinal inflammation of broilers caused by Clostridium perfringens and coccidia, and can be widely used in the field of poultry breeding.

Description

technical field [0001] The invention relates to the technical field of functional microorganism screening and application, in particular to a Bacillus subtilis for preventing and controlling fowl enteritis and its application. Background technique [0002] Intestinal health is an important guarantee for efficient poultry production, but under the dual pressures of increasingly serious bacterial resistance and antibiotic-free breeding, the intestinal problems of poultry are more prominent. Especially in the period of transformation and upgrading of my country's poultry breeding industry, the level of feeding management and disease prevention and control lags behind the process of large-scale and intensive processes, and the incidence of intestinal diseases in poultry farms remains high. The pathogenic bacteria that endanger the intestinal health of poultry mainly include Escherichia coli, Salmonella, Staphylococcus, Campylobacter jejuni, Clostridium perfringens, etc. The acut...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K35/742A61K35/744A61K35/747A61P1/00A61P31/04A61P33/02A23K10/18A23K50/75C12R1/125
CPCC12N1/20A61K35/742A61K35/744A61K35/747A61P1/00A61P31/04A61P33/02A23K10/18A23K50/75A23V2400/125A23V2400/175A23V2400/169A61K2300/00
Inventor 彭虹旎凌红丽李宁周英俊孙文丽梁莉牛庆奎
Owner 山东康地恩生物科技有限公司
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